Method of increasing melatonin synthesis to treat insomnia, ameliorate sleep disturbance and regulate sleep

ABSTRACT

A method for at least one of increasing the synthesis of melatonin and promoting the secretion of melatonin is provided, wherein the method comprises administering to a subject in need an effective amount of a compound of formula (I): 
                         
A method for at least one of treating diseases related to neurometabolic disorder, preventing diseases related to neurometabolic disorder, regulating appetite, regulating mood, regulating vasoconstriction, regulating hemostatic function and regulating immune response is also provided, wherein the method comprises administering to a subject in need an effective amount of a compound of formula (I) as described above.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.16/185,429, filed on Nov. 9, 2018, which claims priority to U.S.Provisional Application Ser. No. 62/590,129, filed on Nov. 22, 2017, andto Taiwan Patent Application No. 107135407, filed on Oct. 8, 2018, thedisclosures of which are incorporated herein in their entirety byreference.

FIELD OF THE INVENTION

The present invention relates to the use of a compound of the followingformula (I) (i.e., hydroxyanigorufone), including the use of compound offormula (I) in increasing the synthesis of melatonin, and/or promotingthe secretion of melatonin:

The present invention also relates to the use of compound of formula (I)in treating diseases related to neurometabolic disorder, preventingdiseases related to neurometabolic disorder, regulating appetite,regulating mood, regulating vasoconstriction, regulating hemostaticfunction, and/or regulating immune response. The present invention alsorelates to the use of compound of formula (I) in increasing expressionof TPH1 gene, increasing expression of DDC gene, increasing expressionof AANAT gene, and/or increasing expression of ASMTL gene.

BACKGROUND OF THE INVENTION

Melatonin, which is a hormone secreted from pineal gland and serves thefunction of adjusting biological clock. People can stay awake during thedaytime because the secretion of melatonin is suppressed by the exposureto light. On the other hand, the secretion of melatonin will increaseduring the nighttime and thus people can fall asleep. The secretion ofmelatonin in the nighttime would be insufficient if one maintains alifestyle of working at night or irregular shifts, being nervous, and/orusing electronics for a long period of time. The insufficient secretionof melatonin will reduce the sleep quality or even cause sleepdisturbances (e.g., insomnia) and/or problems (e.g., feeling tired inthe daytime, messing up the biological clock, decreasing the immunity),and thus seriously affect the daily life.

Medicines generally used in clinic for treating sleep disturbanceinclude sedative-hypnotic drugs (e.g., benzodiazepine and barbiturate)and antidepressants (e.g., clomipramine and imipramine). However, theafore-mentioned medicines are prone to cause addiction and may have sideeffects such as hypersomnia, nausea, headache, vomiting,gastrointestinal discomfort, memory impairment, rebound insomnia,unconsciousness, ataxia, dyspnea, and/or somnambulism. Currently,melatonin being extracted from the pineal glands of cows and/or beingchemically synthesized is also commercially available, while suchmelatonin is strictly controlled or forbidden in the countries that haveconcerns about mad cow disease and/or the uncertain safe dose ofmelatonin. Therefore, there is necessity and urgency for continuouslydeveloping a medicine or method for treating insomnia and/orameliorating sleep disturbances effectively without causing addictionsand side effects.

It was revealed that the expression of melatonin is regulated by genessuch as TPH1 gene, DDC gene, AANAT gene and ASMTL gene, wherein anincreased expression of TPH1 gene, DDC gene, AANAT gene, and/or ASMTLgene is helpful for increasing the synthesis of melatonin, and promotingthe secretion of melatonin, thereby achieves the effects of treatinginsomnia, ameliorating sleep disturbance, and regulating sleep.

Inventors of the present invention discovered that a compound of thefollowing formula (I) (i.e., hydroxyanigorufone) is effective inincreasing the expression of genes such as TPH1 gene, DDC gene, AANATgene and ASMTL gene, and thus, can be used for increasing the synthesisof melatonin and/or promoting the secretion of melatonin, therebyachieves the effects of treating insomnia, ameliorating sleepdisturbances, and regulating sleep. Compound of formula (I) is alsoeffective in treating, preventing or regulating diseases orphysiological functions related to the above genes:

SUMMARY OF THE INVENTION

An objective of the present invention is to provide a use of a compoundof the following formula (I) (i.e., hydroxyanigorufone) in themanufacture of a pharmaceutical composition, wherein the pharmaceuticalcomposition is for increasing the synthesis of melatonin and/orpromoting the secretion of melatonin:

Preferably, the pharmaceutical composition is provided in a form fororal administration, transdermal administration, intravenous injection,or subcutaneous injection. Preferably, the pharmaceutical composition isfor treating insomnia.

Another objective of the present invention is to provide a use of theaforesaid compound of formula (I) in the manufacture of a pharmaceuticalcomposition, wherein the pharmaceutical composition is for at least oneof treating diseases related to neurometabolic disorder, and preventingdiseases related to neurometabolic disorder. Preferably, thepharmaceutical composition is provided in a form for oraladministration, transdermal administration, intravenous injection, orsubcutaneous injection.

Still another objective of the present invention is to provide a use ofthe aforesaid compound of formula (I) in at least one of amelioratingsleep disturbance, regulating appetite, regulating sleep, regulatingmood, regulating vasoconstriction, regulating hemostatic function, andregulating immune response. Preferably, the compound of formula (I) isused for ameliorating sleep disturbance and/or regulating sleep, and istaken through oral, transdermal, or injection route. More preferably,the compound of formula (I) can be used in a form of food composition,wherein the food composition is a health food, a dietary supplement, afunctional food, a nutritional supplement food, or a special nutritionalfood.

Yet another objective of the present invention is to provide a use ofthe aforesaid compound of formula (I) in the manufacture of apharmaceutical composition, wherein the pharmaceutical composition isused for at least one of increasing expression of TPH1 gene, increasingexpression of DDC gene, increasing expression of AANAT gene, andincreasing expression of ASMTL gene. Preferably, the pharmaceuticalcomposition is provided in a form for oral administration, transdermaladministration, intravenous injection, or subcutaneous injection.

Yet another objective of the present invention is to provide a methodfor at least one of increasing the synthesis of melatonin and promotingthe secretion of melatonin, comprising administering to a subject inneed an effective amount of a compound of the following formula (I):

Preferably, the method of the present invention is for treatinginsomnia, ameliorating sleep disturbance, and regulating sleep. In themethod according to the present invention, the compound of formula (I)can be administered as the pharmaceutical composition described above.

Yet another objective of the present invention is to provide a methodfor at least one of treating diseases related to neurometabolicdisorder, preventing diseases related to neurometabolic disorder,regulating appetite, regulating mood, regulating vasoconstriction,regulating hemostatic function and regulating immune response,comprising administering to a subject in need an effective amount of theaforesaid compound of formula (I). In the method according to thepresent invention, the compound of formula (I) can be administered asthe pharmaceutical composition or the food composition described above

Yet another objective of the present invention is to provide a methodfor at least one of increasing expression of TPH1 gene, increasingexpression of DDC gene, increasing expression of AANAT gene andincreasing expression of ASMTL gene, comprising administering to asubject in need an effective amount of the aforesaid compound of formula(I). In the method according to the present invention, the compound offormula (I) can be administered as the pharmaceutical compositiondescribed above.

The detailed technology and preferred embodiments implemented for thepresent invention are described in the following paragraphs accompanyingthe appended drawings for people skilled in this field to wellappreciate the features of the claimed invention.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the effect of compound of formula (I) of the presentinvention on increasing the expressions of TPH1 gene, DDC gene, AANATgene and ASMTL gene, wherein cells in the “control group” were culturedin a medium free of the aforesaid compound of formula (I) for 48 hours,and those in the “experimental group” were cultured in a mediumcontaining the aforesaid compound of formula (I) for 48 hours.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The following will describe some of the embodiments of the presentinvention in detail. However, without departing from the spirit of thepresent invention, the present invention may be embodied in variousembodiments and should not be limited to the embodiments described inthe specification or defined in the appended claims.

Unless otherwise indicated herein, the expressions “a,” “an,” “the,” orthe like recited in the specification of the present invention(especially in the claims) are intended to include both the singular andplural forms. The term “prevent” or “preventing” recited in thisspecification refers to inhibiting or preventing a particular conditionof illness from breaking out, or maintaining good health in a sensitivesubject to tolerate diseases. The term “treat” or “treating” recited inthis specification should not be construed as treating a subject untilthe subject is completely recovered, but should include maintaining theprogression or symptoms of the diseases in a substantially static state,increasing the recovery rate of a subject, alleviating the severity of aparticular condition of illness, or increasing the life quality ofpatients. The term “regulate” or “regulating” recited in thisspecification refers to upregulating (includes inducing, stimulating,and enhancing) or downregulating (includes inhibiting and weakening) thephysiological functions in a subject toward a normal state. The term“subject” recited in this specification refers to a mammalian, includinghuman and non-human animals.

It is known that TPH1 gene is mainly involved in the synthesis ofserotonin, which is related to physiological functions such as appetite,sleep, mood, vasoconstriction, hemostatic function and immune response.Accordingly, the decreased expression of TPH1 gene will affect the abovephysiological functions related to serotonin, and these can be noted insuch as “Analysis of tryptophan hydroxylase I and II mRNA expression inthe human brain: a post-mortem study. J Psychiatr Res. 2007January-February; 41(1-2):168-73,” “Role of Serotonin in the ImmuneSystem and in Neuroimmune Interactions. BRAIN, BEHAVIOR, AND IMMUNITY12, 249-271 (1998),” “Serotonin and sleep. Sleep Medicine Reviews, Vol.6, No. 1, pp 57-69, 2002,” “Effects of Serotonin on Intracellular pH andContraction in Vascular Smooth Muscle. Circulation Research Vol 71,1992, 1294-1304,” “Serotonin and Appetite Regulation. CNS Drugs 1998June; 9 (6): 473-495,” “Serotonin and emotion, learning and memory. Rev.Neurosci. 2012; 23(5-6): 543-553,” and “The role of serotonin inhaemostasis. Hamostaseologie. 2009 November; 29(4):356-9,” which areentirely incorporated hereinto by reference. Therefore, if theexpression of TPH1 gene can be increased, the above physiologicalfunctions related to serotonin can be regulated.

It is known that DDC gene is involved in the synthesis of melatonin, andthe deficiency of DDC gene will also lead to neurometabolic disorder,and these can be noted in such as “A comprehensive picture of themutations associated with aromatic amino acid decarboxylase deficiency:from molecular mechanisms to therapy implications. Hum Mol Genet. 2014Oct. 15; 23(20):5429-40,” which is entirely incorporated hereinto byreference. Therefore, if the expression of DDC gene can be increased,the diseases related to neurometabolic disorder can be treated orprevented, and the physiological functions related to the synthesis ofmelatonin can be regulated.

It is known that AANAT gene plays an important role in the synthesis ofmelatonin, and these can be noted in such as “Diurnal variation in mRNAencoding serotonin N-acetyltransferase in pineal gland. Nature. 1995Dec. 21-28; 378(6559):783-5,” and “Role of a pineal cAMP-operatedarylalkylamine N-acetyltransferasey14-3-3-binding switch in melatoninsynthesis. PNAS Jul. 3, 2001. 98 (14) 8083-8088,” which are entirelyincorporated hereinto by reference. Therefore, if the expression ofAANAT gene can be increased, the physiological functions related to thesynthesis of melatonin can be regulated.

It is known that ASMTL gene is the key gene being responsible for thelatest catalytic step of the synthesis pathway of melatonin, and thesecan be noted in such as “Melatonin synthesis: AcetylserotoninOmethyltransferase (ASMT) is strongly expressed in a subpopulation ofpinealocytes in the male rat pineal gland. Endocrinology. 2016 May;157(5):2028-40,” which is entirely incorporated hereinto by reference.Therefore, if the expression of ASMTL gene can be increased, thephysiological functions related to the synthesis of melatonin can beregulated.

Inventors of the present invention incidentally discovered that thecompound of the following formula (I) could effectively increase theexpressions of TPH1 gene, DDC gene, AANAT gene and ASMTL gene:

Therefore, the present invention relates to the use of compound offormula (I), including: using the compound of formula (I) inameliorating sleep disturbance, regulating appetite, regulating sleep,regulating mood, regulating vasoconstriction, regulating hemostaticfunction and/or regulating immune response, using the compound offormula (I) in manufacturing a pharmaceutical composition, and providinga method of administering to a subject in need an effective amount ofthe compound of formula (I). The pharmaceutical composition and methodprovided in accordance with the present invention are also forincreasing expression of TPH1 gene, DDC gene, AANAT gene and/or ASMTLgene.

The compound of formula (I) adopt in accordance with the presentinvention could be obtained from any suitable source. For example, thecompound of formula (I) could be purchased from the market, be purifiedand isolated from the extract of plant(s), or be obtained by a chemicalsynthesis. For example, the compound of formula (I) could be purchasedfrom the ChemFaces company (Rainbow Biotechnology CO., LTD).

The pharmaceutical composition provided in accordance with the presentinvention could be administered to a subject in need systemically ortopically, and could be delivered by various drug delivery systems(DDSs), such as oral drug delivery system, transdermal drug deliverysystem, injectable drug delivery system, etc. For example, to enhancebioavailability, control drug release speed, target the lesion preciselyand reduce side effects, the pharmaceutical composition could bedelivered by a liposome, a microcapsule, nanoparticles, microneedles,but is not limited thereby.

Depending on the desired purpose(s), the pharmaceutical composition ofthe present invention could be provided in any suitable form withoutparticular limitations. For example, the pharmaceutical compositioncould be provided in a form suitable for administering to a subject inneed by an oral or parenteral (such as transdermal, intravenousinjection, and subcutaneous injection) route, but is not limitedthereby. Depending on the form and purpose(s), a suitable carrier couldbe chosen and used to provide the pharmaceutical composition. Examplesof the carrier include excipients, diluents, auxiliaries, stabilizers,absorbent retarders, disintegrating agents, hydrotropic agents,emulsifiers, antioxidants, adhesives, binders, tackifiers, dispersants,suspending agents, lubricants, hygroscopic agents, etc.

As a form for oral administration, the pharmaceutical composition of thepresent invention can comprise any pharmaceutically acceptable carrierthat will not adversely affect the desired effects of the activeingredient (i.e., the compound of formula (I)). Examples of the suitablecarrier include, but are not limited to, water, saline, dextrose,glycerol, ethanol or its analogs, cellulose, starch, sugar bentonite,and combinations thereof. The pharmaceutical composition could beprovided by any suitable method in any suitable form for oraladministration, such as in the form of a tablet (e.g., dragee), a pill,a capsule, a granule, a pulvis, a fluidextract, a solution, syrup, asuspension, or a tincture, but is not limited thereby.

As a form for transdermal administration, the pharmaceutical compositionof the present invention could be provided in a form of a patch, anemulsion, a cream, a gel (such as a hydrogel), a paste (such as adispersing paste, an ointment), a spray, or a solution (such as asuspension) for external use, but is not limited thereby.

As for the form of injection or drip suitable for subcutaneous orintravenous injection administration, the pharmaceutical composition maycomprise one or more ingredient(s), such as an isotonic solution, asalt-buffered saline (e.g., phosphate-buffered saline orcitrate-buffered saline), a hydrotropic agent, an emulsifier, 5% sugarsolution, and other carriers to provide the pharmaceutical compositionas an intravenous infusion, an emulsified intravenous infusion, a powderfor injection, a suspension for injection, or a powder suspension forinjection. Alternatively, the pharmaceutical composition may be preparedas a pre-injection solid. The pre-injection solid can be provided in aform which is soluble in other solutions or suspensions, or in anemulsifiable form. A desired injection is provided by dissolving thepre-injection solid in other solutions or suspensions or emulsifying itprior to being administered to a subject in need.

Depending on the needs, age, body weight and health conditions of thesubject, the pharmaceutical composition provided in accordance with thepresent invention could be administered at various administrationfrequencies, such as once a day, multiple times a day, or once every fewdays, etc. In addition, the amount of compound of formula (I) in thepharmaceutical composition could be adjusted depending on therequirements of practical application. Optionally, the pharmaceuticalcomposition provided in accordance with the present invention couldfurther comprise one or more other active ingredients (e.g., hypnoticdrugs, antidepressants, and melatonin), or to be used in combinationwith a medicament comprising one or more other active ingredients, tofurther enhance the effects of the pharmaceutical composition, or toincrease the application flexibility and adaptability of the preparationthus provided, as long as the other active ingredients will notadversely affect the desired effects of the active ingredients of thepresent invention (i.e., the compound of formula (I)).

Optionally, the pharmaceutical composition and the food compositionprovided in accordance with the present invention could further comprisea suitable amount of additives, such as a flavoring agent, a toner, or acoloring agent for enhancing the palatability and the visual perceptionof the pharmaceutical composition or the food composition, and/or abuffer, a conservative, a preservative, an antibacterial agent, or anantifungal agent for improving the stability and storability of thepharmaceutical composition or the food composition.

The food composition provided in accordance with the present inventioncan be a health food, a dietary supplement, a functional food, anutritional supplement food or a special nutritional food, and can bemanufactured as dairy products, meat products, breadstuff, pasta,cookies, troche, capsule, fruit juices, teas, sport beverages,nutritional beverages, etc., but is not limited thereby. Preferably thefood composition provided in accordance with the present invention is ahealth food.

Depending on the age, body weight and healthy conditions of the subject,the health food, dietary supplement, functional food, nutritionalsupplement food and special nutritional food provided in accordance withthe present invention could be taken at various frequencies, such asonce a day, several times a day or once every few days, etc. The amountof the compound of formula (I) in the health food, dietary supplement,functional food, nutritional supplement food and special nutritionalfood provided in accordance with the present invention could beadjusted, preferably to the amount that it should be taken daily,depending on the specific population.

The recommended daily dosage, use standards and use conditions for aspecific population (e.g., patient with insomnia, patient withdepression, and pregnant woman), or the recommendations for a use incombination with another food product or medicament can be indicated onthe exterior package of the health food, dietary supplement, functionalfood, nutritional supplement food and/or special nutritional foodprovided in accordance with the present invention. Thus, it is suitablefor the user to take the health food, dietary supplement, functionalfood, nutritional supplement food and/or special nutritional food byhim- or herself safely and securely without the instructions of adoctor, pharmacist, or related executive.

The present invention also provides a method for increasing thesynthesis of melatonin and/or promoting the secretion of melatonin,comprising administering to a subject in need an effective amount of theaforesaid compound of formula (I), wherein the term “a subject in need”refers to a subject having an insufficient synthesis and/or secretion ofmelatonin. The present invention also provides a method for treatingdiseases related to neurometabolic disorder, preventing diseases relatedto neurometabolic disorder, regulating appetite, regulating mood,regulating vasoconstriction, regulating hemostatic function and/orregulating immune response, comprising administering to a subject inneed an effective amount of the aforesaid compound of formula (I),wherein the term “a subject in need” refers to a subject suffering fromneurometabolic disorder-related diseases, an increased appetite, a poorappetite, a mood disorder, vasoconstrictor abnormalities, hemostaticabnormalities and/or an abnormal immune response. In the above method,the compound of formula (I) could be administered to the subject in needas a pharmaceutical composition or a food composition as describedabove, and the administration type, administration route, administrationform, administration frequency and uses of the pharmaceuticalcomposition and the food composition are also all in line with the abovedescriptions.

The present invention also provides a method for increasing expressionof TPH1 gene, increasing expression of DDC gene, increasing expressionof AANAT gene and/or increasing expression of ASMTL gene, comprisingadministering to a subject in need an effective amount of the aforesaidcompound of formula (I), wherein the term “a subject in need” refers toa subject whose TPH1 gene, DDC gene, AANAT gene and/or ASMTL gene isdeleted, mutated, or low-expressed. In this method, the compound offormula (I) could be administered to the subject in need as apharmaceutical composition described above and the administration type,administration route, administration form, administration frequency anduses of the pharmaceutical composition are also all in line with theabove descriptions.

The present invention will be further illustrated in detail withspecific examples as follows. However, the following examples areprovided only for illustrating the present invention and the scope ofthe present invention is not limited thereby. The scope of the presentinvention will be indicated in the appended claims.

EXAMPLES

The sources of materials used in the following Examples are as follows:

-   1. Compound of formula (I) of the present invention: purchased from    ChemFaces company (Rainbow Biotechnology CO., LTD), product number:    CFN96877, CAS number: 56252-02-9.-   2. SHSYSY cell: purchased from ATCC, product number: CRL-2266.-   3. DMEM medium: containing 90% DMEM, 10% FBS and 1%    Penicillin/Streptomycin, wherein DMEM was purchased from Gibco    (product number: 12100-046), FBS was purchased from Gibco (product    number: 10437-028) and Penicillin/Streptomycin was purchased from    Gibco.-   4. RNA Extraction Kit: purchased from Geneaid.-   5. Reverse transcriptase (SuperScript® III Reverse Transcriptase):    purchased from Invitrogen company.-   6. KAPA SYBR FAST qPCR kit: purchased from KAPA Biosystems company.    Step One Plus system: purchased from ABI company.

Example 1

SHSY5Y cells were seeded in a 6-well plate (1×10⁵ cells/well) and thencultured for 24 hours. Thereafter, cells were divided into the controlgroup and experimental group and were separately cultured in thefollowing medium for 48 hours:

-   1. Control group: DMEM medium;-   2. Experimental group: a medium the same as that of the control    group, but additionally added with 0.0625 μg/ml of the compound of    formula (I).

Thereafter, cells of the above groups were harvested and subjected to anRNA extraction with an RNA Extract Kit. The RNA thus obtained was thentranscribed into cDNA by using a Reverse Transcriptase. Thereafter, thecDNA was subjected to a quantitative polymerase chain reaction (qPCR) byusing a Step One Plus system and a KAPA SYBR FAST qPCR kit to determinethe expression levels of genes related to melatonin (including TPH1,DDC, AANAT and ASMTL) in the cells of each group.

The above experimental steps were repeated three times, and then theresults of the three experiments were averaged. Thereafter, the resultof the control group was used as a basis (i.e., the gene expressionlevel of the control group was set as 1-fold) to calculate the relativegene expression level of the experimental group. The results are shownin FIG. 1.

As shown in FIG. 1, as compared to the control group, the expressionlevels of TPH1, DDC, AANAT and ASMTL genes in the cells of theexperimental group all significantly increased. These results indicatethat the compound of formula (I) of the present invention is effectivein increasing the synthesis of melatonin and promoting the secretion ofmelatonin. Hence, the compound of formula (I) of the present inventioncan be used for treating insomnia, ameliorating sleep disturbances,treating diseases related to neurometabolic disorder, preventingdiseases related to neurometabolic disorder, regulating appetite,regulating sleep, regulating mood, regulating vasoconstriction,regulating hemostatic function and regulating immune response,especially for treating insomnia, ameliorating sleep disturbances, andregulating sleep.

What is claimed is:
 1. A method for increasing synthesis of melatonin,comprising administering to a subject in need a composition, wherein thecomposition consists of (a) a compound of formula (I) and (b) at leastone of a pharmaceutical acceptable carrier, an additive, a buffer, and apreservative:


2. The method as claimed in claim 1, wherein the composition isadministered to the subject by at least one of oral administration,transdermal administration, intravenous injection, and subcutaneousinjection.
 3. The method as claimed in claim 1, which results in atleast one of treating insomnia, ameliorating sleep disturbance andregulating sleep.
 4. The method as claimed in claim 3, which is fortreating insomnia.
 5. The method as claimed in claim 3, which is for atleast one of ameliorating sleep disturbance and regulating sleep.
 6. Themethod as claimed in claim 4, wherein the composition is administered tothe subject by at least one of oral administration, transdermaladministration, intravenous injection, and subcutaneous injection. 7.The method as claimed in claim 5, wherein the composition isadministered to the subject by at least one of oral administration andtransdermal administration.